Cheng Dali, Guo Zhiqiang, Zhang Shulan
Department of Obstetrics and Gynaecology, Shengjing Hospital of China Medical University, Shenyang, China.
Contemp Oncol (Pozn). 2015;19(1):36-42. doi: 10.5114/wo.2015.50011. Epub 2015 Mar 26.
In the absence of effective therapeutic strategies, cervical carcinoma continues to be second on the list of mortality rates of malignant tumours found in women. We investigated the effects of β -sitosterol, a natural product isolated from traditional Chinese herbs, on Caski and HeLa cervical carcinoma cells.
Morphological changes were examined by light microscopy. Ultrastructures of Caski and HeLa cells treated with 20 µmol/l β-sitosterol were documented by scanning electron microscopy and transmission electron microscopy. Changes in mRNA and protein expression were quantified respectively using Real-Time qPCR and western blot methods.
Treatment of Caski and HeLa cells with β-sitosterol resulted in reduced expression of PCNA, indicative of an inhibitory effect on cell proliferation. This was associated with increased p53 mRNA levels and decreased amounts of HPV E6 transcripts. Expression of p53 and HPV E6 proteins followed a similar trend as that observed for the corresponding transcripts. Caski and HeLa cells treated with β-sitosterol exhibited loss of cell surface microvilli, increased electron density of cell membrane, and decreased organelles.
In conclusion, treatment of Caski and HeLa cells with β-sitosterol significantly suppressed the expression of genes involved in cell proliferation and oncogenic transformation, while augmenting the expression of genes involved in apoptosis and tumour suppression. Ultrastuctural characterisation of Caski and HeLa cells treated with β-sitosterol further confirmed the anti-proliferative and anti-cancer activity of this natural product isolated from traditional Chinese herbs.
在缺乏有效治疗策略的情况下,宫颈癌在女性恶性肿瘤死亡率排行榜上仍位居第二。我们研究了从传统中药中分离出的天然产物β-谷甾醇对Caski和HeLa宫颈癌细胞的影响。
通过光学显微镜检查形态学变化。用扫描电子显微镜和透射电子显微镜记录用20μmol/lβ-谷甾醇处理的Caski和HeLa细胞的超微结构。分别使用实时定量PCR和蛋白质印迹法对mRNA和蛋白质表达的变化进行定量。
用β-谷甾醇处理Caski和HeLa细胞导致增殖细胞核抗原(PCNA)表达降低,表明对细胞增殖有抑制作用。这与p53 mRNA水平升高和人乳头瘤病毒E6转录本数量减少有关。p53和人乳头瘤病毒E6蛋白的表达与相应转录本的表达趋势相似。用β-谷甾醇处理的Caski和HeLa细胞表现出细胞表面微绒毛丧失、细胞膜电子密度增加和细胞器减少。
总之,用β-谷甾醇处理Caski和HeLa细胞可显著抑制参与细胞增殖和致癌转化的基因表达,同时增强参与细胞凋亡和肿瘤抑制的基因表达。对用β-谷甾醇处理的Caski和HeLa细胞的超微结构表征进一步证实了这种从传统中药中分离出的天然产物的抗增殖和抗癌活性。